Polypeptide cross-linked protein molecularly imprinted polymer and its preparation method and application
A molecular imprinting and polymer technology, applied in chemical instruments and methods, peptides, specific peptides, etc., can solve problems such as poor imprinting effect and difficult template elution
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0029] The invention provides a method for preparing a polypeptide crosslinked protein molecularly imprinted polymer, comprising the following steps: 1) mixing and dissolving a main monomer, a functional monomer, a polypeptide crosslinking agent and a template protein in a water-soluble solvent to obtain a mixed solution; 2) adding an initiator to the mixed solution described in step 1), and polymerizing under the condition that the polypeptide cross-linking agent exists in a helix conformation to obtain a polymer; 3) under the condition that the polypeptide cross-linking agent exists in a coil conformation, Perform template elution on the polymer described in step 2) to obtain a protein molecularly imprinted polymer cross-linked by the polypeptide; the polypeptide cross-linking agent has double bonds capable of polymerizing at both ends of the amino acid sequence, and can generate helix-coil Conformational switched peptides.
[0030]In the present invention, the polypeptide c...
Embodiment 1
[0042] Synthesis of imprinted polymers:
[0043] 200 mg of N-isopropylacrylamide, 5 mg of acrylamide, 4 μL of methacrylic acid, 115 mg of polypeptide cross-linker and 100 mg of template protein lysozyme were dissolved in 2 mL of 20 mM pH5.5 phosphate buffer solution. After thorough mixing, 50 μL of 10% APS and 5 μL TEMED were added to initiate polymerization, and reacted at 37° C. for 24 hours to obtain the imprinted polymer MIP. The non-imprinted polymer NIP was prepared in the same way as the imprinted polymer, except that the template protein lysozyme was not added. The structure of the polypeptide cross-linking agent is shown in formula I:
[0044]
[0045] Elution of template protein:
[0046] Protein elution was carried out at 37° C. with 20 mM pH7.4 phosphate buffer solution (containing 0.154 M NaCl to simulate physiological ionic strength). Measure the absorbance value of the eluate, use a UV spectrophotometer to draw a standard curve of absorbance-concentration ...
Embodiment 2
[0060] Synthesis of imprinted polymers:
[0061] Dissolve 200 mg N-isopropylacrylamide, 2.5 mg acrylamide, 8 μL dimethylaminoethyl methacrylate, 115 mg polypeptide cross-linker and 100 mg template protein bovine serum albumin (BSA) in 2 mL 20 mM pH5.5 phosphate buffer solution. After mixing well, add initiator 50 μL 10% APS and 5 μL TEMED, react at 37° C. for 24 h. Obtain the imprinted polymer MIP. The non-imprinted polymer NIP was prepared in the same way as the imprinted polymer, except that the template protein BSA was not added. The structure of the polypeptide cross-linking agent is the same as in Example 1.
[0062] Elution of template protein:
[0063] Protein was eluted with 20mM pH7.4 phosphate buffer solution (containing 0.154M NaCl to simulate physiological ionic strength) at 37°C, and the four elution rates were 95.24%, 2.05%, 0.24%, and 0%, respectively. The total elution rate was 97.51%. Finally, the imprinted polymer was washed with deionized water to remo...
PUM
Login to view more Abstract
Description
Claims
Application Information
Login to view more - R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap